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Thromb Haemost 1993; 70(04): 654-658
DOI: 10.1055/s-0038-1649644
Original Article
Platelets
Schattauer GmbH Stuttgart

Complexes of Nitric Oxide with Nucleophiles as Agents for the Controlled Biological Release of Nitric Oxide: Antiplatelet Effect

Jean G Diodati
1   The Cardiology Branch, National Heart, Lung, and Blood Institute, Bethesda, Maryland, USA
,
Arshed A Quyyumi
1   The Cardiology Branch, National Heart, Lung, and Blood Institute, Bethesda, Maryland, USA
,
Nabeen Hussain
1   The Cardiology Branch, National Heart, Lung, and Blood Institute, Bethesda, Maryland, USA
,
Larry K Keefer
2   Chemistry Section, Laboratory of Comparative Carcinogenesis, National Cancer Institute, Frederick Cancer Research and Development Center, Frederick, Maryland, USA
› Author Affiliations
Further Information

Publication History

Received 21 September 1992

Accepted after revision 24 May 1993

Publication Date:
05 July 2018 (online)

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Summary

Nitric oxide (NO) inhibits platelet aggregation. Accordingly, we hypothesized that complexes of diethylamine and spermine with NO (DEA/NO and SPER/NO, respectively), two vasodilators previously shown to release NO spontaneously in aqueous solution, may also be useful antiplatelet agents. Platelet aggregation was measured in whole blood or platelet-rich plasma by impedance aggregometry after addition of collagen. In whole blood, the dose response curve for DEA/NO added 1 min before collagen was similar to that for aspirin (60% inhibition at 10-4 M), while SPER/NO and sodium nitroprusside were less potent by an order of magnitude. In platelet-rich plasma, 10-6 M DEA/NO caused 60% inhibition, while SPER/NO and sodium nitroprusside were as active only at 10-5 M; aspirin’s potency was unchanged from that in whole blood. In vivo, DEA/NO and sodium nitroprusside produced significant platelet inhibition 1 min after intravenous injection in the rabbit at 50 nmol/kg. Similar in vivo platelet inhibition was observed with SPER/NO and aspirin, but only at higher dose. The effects of DEA/NO and sodium nitroprusside were transient, lasting less than 30 min after treatment, while the activity of SPER/NO and aspirin was sustained throughout the 30 min experiment. The magnitude and duration of the antiplatelet effects of DEA/NO and SPER/NO correlate with the rates at which they release nitric oxide spontaneously in aqueous solution. Thus, NO/nucleophile complexes merit further exploration both as research tools and as potential antiplatelet agents.

 

  • References

  • 1 Furchgott RF, Vanhoutte PM. Endothelium-derived relaxing and contracting factors. FASEB J 1989; 3: 2007-2018
    CrossrefPubMedGoogle Scholar
  • 2 Lüscher TF, Vanhoutte PM. The Endothelium: Modulator of Car-diovascular Function. Boca Raton, FL: CRC Press; 1990
    PubMedGoogle Scholar
  • 3 Moncada S, Palmer RMJ, Higgs EA. Nitric oxide: Physiology, pathophysiology, and pharmacology. Pharmacol Rev 1991; 43: 109-142
    PubMedGoogle Scholar
  • 4 Myers PR, Minor Jr RL, Guerra Jr R, Bates JN, Harrison DG. Vasorelaxant properties of the endothelium-derived relaxing factor more closely resemble S-nitrosocysteine than nitric oxide. Nature 1990; 345: 161-163
    CrossrefPubMedGoogle Scholar
  • 5 Ignarro LJ. Biological actions and properties of endothelium-derived nitric oxide formed and released from artery and vein. Circ Res 1989; 65: 1-21
    CrossrefPubMedGoogle Scholar
  • 6 Marletta MA, Tayeh MA, Hevel JM. Unraveling the biological significance of nitric oxide. BioFactors 1990; 2: 219-225
    PubMedGoogle Scholar
  • 7 Stamler JS, Simon DI, Osborne JA, Mullins ME, Jaraki O, Michel T, Singel DJ, Loscalzo J. S-Nitrosylation of proteins with nitric oxide: Synthesis and characterization of biologically active compounds. Proc Natl Acad Sci USA 1992; 89: 444-448
    CrossrefPubMedGoogle Scholar
  • 8 Bassenge E, Busse R. Endothelial modulation of coronary tone. Prog Cardiovasc Dis 1988; 30: 349-380
    CrossrefPubMedGoogle Scholar
  • 9 Furchgott RF, Zawadzki JV. The obligatory role of endothelial cells in the relaxation of arterial smooth muscle by acetylcholine. Nature 1980; 288: 373-376
    CrossrefPubMedGoogle Scholar
  • 10 Furchgott RF. Role of endothelium in responses of vascular smooth muscle. Circ Res 1983; 53: 557-573
    CrossrefPubMedGoogle Scholar
  • 11 Ignarro LJ, Buga GM, Wood KS, Byrns RE, Chaudhuri G. Endothelium-derived relaxing factor produced and released from artery and vein is nitric oxide. Proc Natl Acad Sci USA 1987; 84: 9265-9269
    CrossrefPubMedGoogle Scholar
  • 12 Palmer RMJ, Ferrige AG, Moncada S. Nitric oxide release accounts for the biological activity of endothelium-derived relaxing factor. Nature 1987; 327: 524-526
    CrossrefPubMedGoogle Scholar
  • 13 de Graaf JC, Banga JD, Moncada S, Palmer RMJ, de Groot PG, Sixma JJ. Nitric oxide functions as an inhibitor of platelet adhesion under flow conditions. Circulation 1992; 85: 2284-2290
    CrossrefPubMedGoogle Scholar
  • 14 Vallance P, Collier J, Moncada S. Effects of endothelium-derived nitric oxide on peripheral arteriolar tone in man. Lancet 1989; ii: 997-1000
    PubMedGoogle Scholar
  • 15 Moncada S, Radomski MW, Palmer RMJ. Endothelium-derived relaxing factor: Identification as nitric oxide and role in the control of vascular tone and platelet function. Biochem Pharmacol 1988; 37: 2495-2501
    CrossrefPubMedGoogle Scholar
  • 16 Radomski MW, Palmer RMJ, Moncada S. An L-arginine/nitric oxide pathway present in human platelets regulates aggregation. Proc Natl Acad Sci USA 1990; 87: 5193-5197
    CrossrefPubMedGoogle Scholar
  • 17 Mollace V, Salvemini D, Vane J. Studies on the importance of the proposed release of nitric oxide from platelets. Thromb Res 1991; 64: 533-542
    CrossrefPubMedGoogle Scholar
  • 18 Waldman SA, Murad F. Cyclic GMP synthesis and function. Pharmacol Rev 1987; 39: 163-196
    PubMedGoogle Scholar
  • 19 Bhardwaj R, Page CP, May GR, Moore PK. Endothelium-derived relaxing factor inhibits platelet aggregation in human whole blood in vitro and in the rat in vivo. Eur J Pharmacol 1988; 157: 83-91
    CrossrefPubMedGoogle Scholar
  • 20 Busse R, Lückhoff A, Bassenge E. Endothelium-derived relaxant factor inhibits platelet activation. Naunyn Schmiedebergs Arch Pharmacol 1987; 336: 566-571
    PubMedGoogle Scholar
  • 21 Azuma H, Ishikawa M, Sekizaki S. Endothelium-dependent inhibition of platelet aggregation. Br J Pharmacol 1986; 88: 411-415
    CrossrefPubMedGoogle Scholar
  • 22 Radomski MW, Jenkins DC, Holmes L, Moncada S. Human colorectal adenocarcinoma cells: Differential nitric oxide synthesis determines their ability to aggregate platelets. Cancer Res 1991; 51: 6073-6078
    PubMedGoogle Scholar
  • 23 Broekman MJ, Eiroa AM, Marcus AJ. Inhibition of human platelet reactivity by endothelium-derived relaxing factor from human umbilical vein endothelial cells in suspension: Blockade of aggregation and secretion by an aspirin-insensitive mechanism. Blood 1991; 78: 1033-1040
    PubMedGoogle Scholar
  • 24 Furlong B, Henderson AH, Lewis MJ, Smith JA. Endothelium-derived relaxing factor inhibits in vitro platelet aggregation. Br J Pharmacol 1987; 90: 687-692
    CrossrefPubMedGoogle Scholar
  • 25 Radomski MW, Palmer RMJ, Moncada S. Comparative pharmacology of endothelium-derived relaxing factor, nitric oxide and pros-tacyclin in platelets. Br J Pharmacol 1987; 92: 181-187
    CrossrefPubMedGoogle Scholar
  • 26 Radomski MW, Palmer RMJ, Moncada S. The anti-aggregating properties of vascular endothelium: Interactions between prostacyclin and nitric oxide. Br J Pharmacol 1987; 92: 639-646
    CrossrefPubMedGoogle Scholar
  • 27 Nishimura H, Rosenblum WI, Nelson GH, Boyton S. Agents that modify EDRF formation alter antiplatelet properties of brain arteriolar endothelium in vivo. Am J Physiol 1991; 261: H15-H21
    PubMedGoogle Scholar
  • 28 Maurice DH, Haslam RJ. Molecular basis of the synergistic inhibition of platelet function by nitrovasodilators and activators of adenylate cyclase: Inhibition of cyclic AMP breakdown by cyclic GMP. Mol Pharmacol 1990; 37: 671-681
    PubMedGoogle Scholar
  • 29 Gerzer R, Drummer C, Karrenbrock B, Heim J-M. Inhibition of platelet activating factor-induced platelet aggregation by mol-sidomine, SIN-1, and nitrates in vitro and ex vivo. J Cardiovasc Pharmacol 1989; 14 (Suppl. 11) S 115-S 119
    CrossrefPubMedGoogle Scholar
  • 30 Mellion BT, Ignarro LJ, Ohlstein EH, Pontecorvo EG, Hyman AL, Kadowitz PJ. Evidence for the inhibitory role of guanosine 3’,5’-monophosphate in ADP-induced human platelet aggregation in the presence of nitric oxide and related vasodilators. Blood 1981; 57: 946-955
    PubMedGoogle Scholar
  • 31 Pohl U, Busse R. EDRF increases cyclic GMP in platelets during passage through the coronary vascular bed. Circ Res 1989; 65: 1798-1803
    CrossrefPubMedGoogle Scholar
  • 32 Shimokawa H, Vanhoutte PM. Hypercholesterolemia causes generalized impairment of endothelium-dependent relaxation to aggregating platelets in porcine arteries. J Am Coll Cardiol 1989; 13: 1402-1408
    CrossrefPubMedGoogle Scholar
  • 33 Drago RS. Reactions of nitrogen(II) oxide. in Colburn CB. (ed.) Free Radicals in Inorganic Chemistry (Advances in Chemistry Series, Number 36). Washington, DC: American Chemical Society; 1962: 143-149
  • 34 Maragos CM, Morley D, Wink DA, Dunams TM, Saavedra JE, Hoffman A, Bove AA, Isaac L, Hrabie JA, Keefer LK. Complexes of NO with nucleophiles as agents for the controlled biological release of nitric oxide. Vasorelaxant effects. J Med Chem 1991; 34: 3242-3247
    CrossrefPubMedGoogle Scholar
  • 35 Cardinal DC, Flower RJ. The electronic aggregometer: A novel device for assessing platelet behavior in blood. J Pharmacol Methods 1980; 3: 135-158
    CrossrefPubMedGoogle Scholar
  • 36 Mackie IJ, Jones R, Machin SJ. Platelet impedance aggregation in whole blood and its inhibition by antiplatelet drugs. J Clin Pathol 1984; 37: 874-878
    CrossrefPubMedGoogle Scholar
  • 37 Galvez A, Badimon L, Badimon J-J, Fuster V. Electrical aggregometry in whole blood from human, pig and rabbit. Thromb Haemostas 1986; 56: 128-132
    PubMedGoogle Scholar
  • 38 Diodati JG, Quyyumi AA, Keefer LK. Complexes of nitric oxide with nucleophiles as agents for the controlled biological release of nitric oxide: Hemodynamic effect in the rabbit. J Cardiovasc Pharmacol 1993; 22: 287-292
    CrossrefPubMedGoogle Scholar
  • 39 Diodati J, Théroux P, Latour J-G, Lacoste L, Lam JYT, Waters D. Effects of nitroglycerin at therapeutic doses on platelet aggregation in unstable angina pectoris and acute myocardial infarction. Am J Cardiol 1990; 66: 683-688
    CrossrefPubMedGoogle Scholar
  • 40 Diodati JG, Cannon III RO, Epstein SE, Quyyumi AA. Increased platelet aggregability across the coronary bed in response to rapid atrial pacing in patients with stable coronary artey disease. Circulation 1992; 86: 1186-1193
    CrossrefPubMedGoogle Scholar
  • 41 Saxon A, Kattlove HE. Platelet inhibition by sodium nitroprusside, a smooth muscle inhibitor. Blood 1976; 47: 957-961
    PubMedGoogle Scholar
  • 42 Mehta J, Mehta P. Comparative effects of nitroprusside and nitrogly-cerin on platelet aggregation in patients with heart failure. J Cardio-vasc Pharmacol 1980; 2: 25-33
    CrossrefPubMedGoogle Scholar
  • 43 Böhme E, Graf H, Schultz G. Effects of sodium nitroprusside and other smooth muscle cell relaxants on cyclic GMP formation in smooth muscle and platelets. Adv Cyclic Nucleotide Res 1978; 9: 131-143
    PubMedGoogle Scholar
  • 44 Levin RI, Weksler BB, Jaffe EA. The interaction of sodium nitro-prusside with human endothelial cells and platelets: Nitroprusside and prostacyclin synergistically inhibit platelet function. Circulation 1982; 66: 1299-1307
    CrossrefPubMedGoogle Scholar
  • 45 Houston DS, Robinson P, Gerrard JM. Inhibition of intravascular platelet aggregation by endothelium-derived relaxing factor: Reversal by red blood cells. Blood 1990; 76: 953-958
    PubMedGoogle Scholar
  • 46 Bates JN, Baker MT, Guerra Jr R, Harrison DG. Nitric oxide generation from nitroprusside by vascular tissue. Evidence that reduction of the nitroprusside anion and cyanide loss are required. Biochem Pharmacol 1991; 42 (Suppl. 00) S 157-S 165
    CrossrefPubMedGoogle Scholar
  • 47 Roth GJ, Stanford N, Majerus PN. Acetylation of prostaglandin synthase by aspirin. Proc Natl Acad Aci USA 1975; 72: 3073-3076
    CrossrefPubMedGoogle Scholar
  • 48 Morley D, Maragos CM, Zhang X-Y, Boignon M, Wink DA, Keefer LK. Mechanism of vascular relaxation induced by the nitric oxide (No)/nucleophile complexes, a new class of based vasodilators. J Cardiovasc Pharmacol 1993; 21: 670-676
    CrossrefPubMedGoogle Scholar
  • 49 Maragos CM, Wang JM, Hrabie JA, Oppenheim JJ, Keefer LK. Nitric oxide/nucleophile coplexes inhibit the in vitro proliferation of A375 melanoma cells via nitric oxide release. Cancer Res 1993; 53: 564-568
    PubMedGoogle Scholar